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塞来昔布通过抑制端粒酶和诱导凋亡对Hep-2细胞产生抗增殖作用。

Antiproliferative effects of celecoxib in Hep-2 cells through telomerase inhibition and induction of apoptosis.

作者信息

Zhao Yong-Qiang, Feng Hui-Wei, Jia Tao, Chen Xue-Mei, Zhang Hui, Xu An-Ting, Zhang Hai-Ling, Fan Xian-Liang

机构信息

Department of Otolaryngology and Head and Neck Surgery, the Second Hospital of Shandong University, Jinan, China E-mail :

出版信息

Asian Pac J Cancer Prev. 2014;15(12):4919-23. doi: 10.7314/apjcp.2014.15.12.4919.

Abstract

BACKGROUND

To investigate the effect of celecoxib on telomerase activity and apoptosis in a human laryngeal squamous carcinoma cell line (Hep-2 cells).

MATERIALS AND METHODS

The growth inhibition rate of Hep-2 cells in vitro was measured by MTT assay, and apoptosis by TUNEL assay and flow cytometry (FCM). The TRAP-ELISA method was used to determine telomerase activity in Hep-2 cells. The mRNA expression of human telomerase RNA component(hTR), human telomerase reverse transcriptase (hTERT) and human telomerase-associated protein(hTEP1) was determined by RT-PCR assay. Expression of Bax and Bcl-2 proteins was assessed by Western blotting.

RESULTS

Celecoxib can inhibit proliferation and induce apoptosis in a dose- and time-dependent manner, repress telomerase activity, decrease hTERT mRNA and Bcl-2 protein expression and increase Bax protein expression, PGE2 had no effect on telomerase.

CONCLUSIONS

Celecoxib had the antiproliferative and pro-apoptotic effect in Hep-2 cells. Apoptosis was accompanied by a decrease in telomerase activity which was directly correlated with hTERT mRNA and up-regulation of Bax/Bcl-2. Bcl-2 may thus play an important role in telomerase activity as well as apoptosis.

摘要

背景

研究塞来昔布对人喉鳞状癌细胞系(Hep-2细胞)端粒酶活性及凋亡的影响。

材料与方法

采用MTT法检测Hep-2细胞的体外生长抑制率,采用TUNEL法和流式细胞术(FCM)检测细胞凋亡。采用TRAP-ELISA法测定Hep-2细胞的端粒酶活性。采用RT-PCR法检测人端粒酶RNA组分(hTR)、人端粒酶逆转录酶(hTERT)及人端粒酶相关蛋白(hTEP1)的mRNA表达。采用蛋白质印迹法检测Bax和Bcl-2蛋白的表达。

结果

塞来昔布能以剂量和时间依赖性方式抑制增殖并诱导凋亡,抑制端粒酶活性,降低hTERT mRNA和Bcl-2蛋白表达,增加Bax蛋白表达,PGE2对端粒酶无影响。

结论

塞来昔布对Hep-2细胞具有抗增殖和促凋亡作用。凋亡伴随着端粒酶活性降低,这与hTERT mRNA及Bax/Bcl-2上调直接相关。因此,Bcl-2可能在端粒酶活性及凋亡中起重要作用。

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